M. tuberculosis (MTb) infects one-third of the world?s population, causes TB disease in 9 million people per year, and is responsible for 1.7 million deaths annually. While effective treatments for TB have been available for over 50 years, a major barrier to its control has remained the inability to reliably diagnose disease in low-resource TB-endemic settings. Most clinical tests require an adequate sample of sputum (mucus from the lungs) which may be difficult to obtain in patients without a cough, in children, and in HIV-infected individuals. Urine offers an attractive method for point-of-care TB diagnostics since it is abundant, simple to collect, and collection does not involve exposure to infectious aerosols. Point-of-care TB diagnostics are urgently needed to decrease disease transmission and improve treatment outcomes. The development of a PCR-based test of TB from urine, and its development as a point-of-care test, could have enormous impact on TB diagnostics. Aim 1. Demonstrate the ability to diagnose pulmonary TB from patient urine. We will evaluate the performance of detection for diagnosing pulmonary TB from urine of patients within seven days of treatment initiation. We will calculate sensitivity in the 30 subjects known to have pulmonary TB based on sputum analysis results, and calculate specificity in 50 healthy controls. Aim 2. Enhance performance by expanding targets to additional regions of the TB genome. Extensive analysis of the sensitivity and specificity of diagnostic targets in TB genomes is lacking. We will analyze TB genomes in the NCBI GenBank to determine inclusiveness and specificity. We will assess common targets in combination with other reported targets to identify a strong set for high accuracy. As these assays are developed, we will re-test urine samples from Aim 1. Aim 3. Demonstrate a point-of-care-compatible device based on quantitative engineering of capture chemistry and conditions. We will quantify performance to allow informed engineering to demonstrate a proof-of-principle device. The chosen demonstration will not be a final device but will demonstrate a proof-of-principle that is aligned with the design goals: rapid processing, self-contained shelf-stable reagents, no lab equipment, low-cost disposable components, simple user steps, target recovery >80% from ~1 mL urine.